32 research outputs found

    Hyphal tip growth and cytoplasmic characters of <i>Conidiobolus coronatus</i> (Zoopagomycota, Entomophthoromycotina)

    No full text
    <p>Characteristics of hyphal structure and growth can provide insights into the mechanisms of polarized growth and support investigations of fungal phylogeny. To assist with the resolution of evolutionary relationships of the zygomycetes, the authors used comparative bioimaging methods (light [LM] and transmission electron [TEM] microscopy) to describe selected subcellular characters of hyphal tips of <i>Conidiobolus coronatus</i>. Growing hyphae of <i>C. coronatus</i> contain Spitzenkörper (Spk). Spk are most commonly present in hyphae of Dikarya (Ascomycota and Basidiomycota) and are rarely reported in zygomycete hyphae, which possess an apical vesicle crescent (AVC). Such findings raise questions regarding the evolution of the Spk and its relationship with the AVC. Descriptions of additional subcellular characters (e.g., mitotic-phase spindle pole bodies, cytoplasmic behavior, organelle structure) are also presented.</p

    Reconstructions of morphogenetic sequences from time lapse movies of the Δ<i>crn-1</i> mutant and the WT strain; the mutant shows a lack of directionality and temporal loss of polarized growth, the green dots show the places of endocytic patches and how they are misplaced in the mutant when the Spk is not present, the small arrows show the shifts between polarized growth and isotropic growth.

    No full text
    <p>Reconstructions of morphogenetic sequences from time lapse movies of the Δ<i>crn-1</i> mutant and the WT strain; the mutant shows a lack of directionality and temporal loss of polarized growth, the green dots show the places of endocytic patches and how they are misplaced in the mutant when the Spk is not present, the small arrows show the shifts between polarized growth and isotropic growth.</p

    Effect of cytoskeleton depolymerization drugs on the localization and integrity of coronin patches.

    No full text
    <p>Hyphae exposed to: (A) the anti-actin drug, 1.0 µg ml<sup>−1</sup> cytochalasin A, (B) 5.0 µg ml<sup>−1</sup> cytochalasin A, and (C) the anti-tubulin drug 2.5 µg ml<sup>−1</sup> benomyl. Scale bar = 5 µm.</p

    Comparison of conidial morphology and size.

    No full text
    <p>Conidiophores of (A) Δ<i>crn-1</i> mutant and (B) WT strain. Composite image of conidia representing the most common shapes in (C) Δ<i>crn-1</i> mutant and (D) WT strain. (E) Relative abundance of spherical and non-spherical conidia in the Δ<i>crn-1</i> mutant and WT strain. (F) Average size of conidia in the Δ<i>crn-1</i> mutant and WT strain. The error bars represent the 95% confidence interval. Scale bar = 5 µm.</p

    Co-expression of coronin with fimbrin, Arp2 and actin.

    No full text
    <p>(A–C) Colocalization of Fimbrin (FIM-GFP) and CRN-1-mChFP. (D–F) Colocalization of Arp2 (ARP-2-GFP) and CRN-1-mChFP. (G–I) Partial colocalization of the actin marker Lifeact-GFP and CRN-1-mChFP. (J–L) Co-expression of CRN-1-mChFP and Lifeact-GFP showing the lack of colocalization between coronin patches and actin cables. are depicted by. (L) Merge, not clear association of crn-1 patches is observed with actin filaments, arrowhead shows colocalization of actin patches with CRN-1-mChFP. The white arrow points a region where there is only labeling with Lifeact-GFP and the blue arrow show the patches where CRN-1-mChFP and Lifeact-GFP colocalized. Note the presence of actin in the Spk but not of patch related ABPs. The red arrows in (K) point the actin cables and the white arrowhead show the colocalization of actin and coronin in the patches subapical collar. Scale bar = 5 µm.</p

    Subapical localization of coronin.

    No full text
    <p>(A) CRN-1-GFP forms a subapical collar along the inner perimeter of the hypha (arrows), (B) FM4-64 staining reveals the position of the Spk (arrowheads), (C) merge of CRN-1-GFP and FM4-64 staining shows the absence of CRN-1-GFP in the Spk, single confocal plane images. (D) 3D reconstruction of merged confocal z-stacks showing CRN-1-GFP and FM4-64 localization, (E) orthogonal view of the 3D reconstruction shown in (D), the yellow line indicates the position within the tip where the cross-section was taken. Scale bars = 5 µm.</p

    Conidial germination.

    No full text
    <p>(A–E) Time series of the Δ<i>crn-1</i> mutant by bright field microscopy and (F–J) WT strain by phase contrast microcopy. Reconstruction of the morphological differences during conidial germination of above sequences over longer time periods (K) Δ<i>crn-1</i> mutant and (L) WT. Time in h:min. Scale bar = 5 µm.</p

    Dynamics of the subapical endocytic collar and Spk behavior in the Δ<i>crn-1</i> mutant.

    No full text
    <p>(A–L) Subapical endocytic collar of fimbrin, (A–D) Fimbrin-GFP, (E–H) FM4-64 staining and (I–L) merge of Fimbrin-GFP and FM4-64. (M–Y) actin distribution shown with (M–P) Lifeact-GFP, (R–T) staining with FM4-64 and (V–Y) merge of Lifeact-GFP and FM4-64. White arrows point to the presence of the Spk. Time in min:sec. Scale bar = 10 µm. Note. The hypha Q–T was exposed to FM4-64 for a longer time prior to the start of the sequence, hence the stronger red signal.</p

    Phenotype of Δ<i>crn-1</i> mutant.

    No full text
    <p><b>Colony morphology of (A–B) Δ</b><b><i>crn-1</i></b><b> mutant and (C–D) WT strain after 24 and 48 h of incubation on VMM at 28°C.</b> Low magnification images of the colony edge of (E) Δ<i>crn-1</i> mutant and (F) WT strain. Phase contrast images of hypha of (G) Δ<i>crn-1</i> mutant and (H) WT strain. SEM images of (I) the meandering phenotype of Δ<i>crn-1</i> mutant hyphae and (J) the straight WT hyphae. TEM images of the subapical region of 6 h-old germlings of (K) Δ<i>crn-1</i> mutant and (l) WT strain. A comparison of the uneven thickness of the cell wall and the ruffled plasma membrane of the mutant (arrow) with the uniform envelope of the WT (arrowhead). Scale bars = (A–D) 2.5 cm, (E–F) 100 µm, (G–J) 10 µm, (K–L) 1.0 µm.</p
    corecore